Lysine acetylation can be defined as the transfer of an acetyl group to the lysine residues in proteins, this process is mediated by acetyltransferases that are divided into several families. Similarly, deacetylation is brought about by several different classes of protein deacetylases. The roles of lysine acetylation in the regulation of gene expression through the reversible modification of core histone tails or transcription factors were well studied. Lysine acetylation has been found to play important roles in various other cellular processes since 1996 in some independent studies. However, until the year 2009, high-throughput mass spectrometry makes it possible to study acetylation in proteomics levels. Lysine acetylation could be found in almost everywhere. Scientists found that lysine acetylation preferentially targets proteins involved in diverse cellular processes, such as chromatin remodeling, cell cycle, nuclear transport, and cellular metabolism. In this review, we will focus on the roles of lysine acetylation on cellular metabolism, and illustrate that reversible acetylation of metabolic enzymes represents a metabolic regulatory mechanism conserved from bacteria to mammals.